Effects Of10-HDA On The Development Of Honeybee (Apis Mellifera Ligustica) Larvae And Analysis Of MRNA Expression And Prokaryotic Expression Of Royalactin In Apis Cerana Cerana

Worker and queen honeybees develop from the identical genome, but there areremarkable morphological, behavioural, and physiological differences. It is well knownthat the developmental plasticity of honeybee results from differential intake of royal jellyat an early larvae stage. For the first3days of life, all larvae were fed with royal jelly.Worker larvae were then switched to a diet of pollen and nectar, whereas queen larvaeremain on a royal-jelly diet throughout their larval development. From this, it can be seenthat royal jelly is the most important element to induce the caste differentiation ofhoneybee. Recently, researches on its components and functions have bee hotspot onapicultural reasearch fields.As the (E)-10-Hydroxy-2-decenoic acid (10-HDA) which can inhibit the histonedeacetylation is a main component of the royal jelly. The newly hatched larvae wereartificially fed with diets containing1.10%,1.70%,2.30%,2.90%of10-HDA, regarding tocontrol (C), treatment1(T1), treatment2(T2), treatment3(T3), respectively, to study itseffects on the larvae cast differentiation. Comparing with the control group, the weight ofthe emerged adults was significantly reduced in the10-HDA-treated group. The expressionlevel of histone deacetylase3(HDAC3) gene was significantly up-regulated in the10-HDA-treated group than in the control group. The expression level of DNAmethyltranferase3(DNMT3) gene was down-regulated at first, but then up-regulated withthe increasing of10-HDA concentration. The results suggested acetylation and methylationmay dynamically work together to regulate the honey bee cast differentiation.This study was also conducted to explore royalactin (also called Major Royal JellyProteins1，MRJP1) gene of Apis cerana cerana. Firstly, the expression level ofAcc-royalactin gene at different developmental stages (newly emerged bee，NEB；nursebee，NB；forager bee，FB) and two kinds of nurse bee (nurse breeding worker larvae，NBWL；nurse breeding queen larvae，NBQL) were detected by qRT-PCR. Secondly, theentire ORF of Acc-royalactin gene was amplified from the heads of8-day-old worker ofApis cerana cerana. The Acc-royalactin gene was sub-cloned into the prokaryoticexpression vector pSumo-Mut for fusion expression in ArcticExpressTM（DE3）. The resultsshowed that the expression levels of Acc-royalactin in NB and FB were significantlyincreased than that in NEB (p<0.05), and there was no significant difference betweenNBWL and NBQL on the expression level of Acc-royalactin. The recombinant plasmidpSumo-Mut/Acc-royalactin was transformed into ArcticExpressTM（DE3） and induced byIPTG, the SDS-PAGE results showed recombinant protein royalactin-Sumo had specific bands...